Compositions and Methods for Inducing Protein Function

Abstract

Provided herein are compositions and methods for inducing protein function. For example, in some embodiments, provided herein are compositions and methods for pharmacological induction of protein function.

1 . A composition, comprising:
 a fusion protein comprising a) a first polypeptide of interest; and b) a first protease and a substrate for said protease.   
     
     
         2 . The composition of  claim 1 , wherein said protease and said substrate are inserted between multiple domains of said polypeptide. 
     
     
         3 . The composition of  claim 1 , wherein said protease and said substrate are inserted within a domain of said polypeptide of interest. 
     
     
         4 . The composition of  claim 1 , wherein said protease and said substrate are inserted between two copies of said polypeptide of interest. 
     
     
         5 . The composition of any one of  claims 1  to  4 , wherein said composition comprises a second fusion protein comprising second polypeptide of interest and a second protease, wherein said second protease is distinct from said first protease. 
     
     
         6 . The composition of  claim 5 , wherein said first protease and said second protease are inhibited by different protease inhibitors. 
     
     
         7 . The composition of any one of  claims 1  to  6 , wherein said first and second proteases are HCV NS3 proteases. 
     
     
         8 . The composition of  claim 7 , wherein said HCV NS3 proteases comprise at least one mutation. 
     
     
         9 . The composition of  claim 8 , wherein said first HCV NS3 protease comprises V36M, T54A, and S122G mutations and said second HCV NS3 protease comprises F43L, Q80K, S122R, and D168Y mutations. 
     
     
         10 . The composition of  claim 9 , wherein said HCV NS3 protease comprising V36M, T54A, and S122G mutations is resistant to telaprevir (TPV) and sensitive to asunaprevir (ASV) and said HCV NS3 protease comprising F43L, Q80K, S122R, and D168Y mutations is resistant to ASV and sensitive to TPV. 
     
     
         11 . A nucleic acid encoding the fusion protein of any one of  claims 1  to  10 . 
     
     
         12 . A cell comprising the nucleic acid of  claim 11  or the fusion protein of any one of  claims 1  to  10 . 
     
     
         13 . The cell of  claim 12 , wherein said nucleic acid is on a chromosome of said cell. 
     
     
         14 . The cell of  claim 12  or  13 , wherein said cell is in an organism. 
     
     
         15 . The cell of  claim 14 , wherein said organism is selected from the group consisting of a microorganism, a non-human animal, and a human. 
     
     
         16 . The composition of any one of  claims 1  to  10 , wherein said protein of interest is selected from the group consisting of a transcription factor, a nuclease enzyme, a protease enzyme, and a metabolic enzyme. 
     
     
         17 . A kit or system, comprising:
 a) the nucleic acid of  claim 11 ; and   b) at least one protease inhibitor.   
     
     
         18 . A method of modulating the activity or function of a polypeptide of interest, comprising:
 contacting the cell of any one of  claims 12  to  15  with a protease inhibitor under conditions such that said polypeptide of interest is active.   
     
     
         19 . The method of  claim 18 , wherein said polypeptide of interest is selected from the group consisting of a transcription factor, a nuclease enzyme, a protease enzyme, and a metabolic enzyme.